Cunnick J, Rider M, Takemoto L J, Takemoto D J
Department of Biochemistry, Kansas State University, Manhattan 66506.
Biochem J. 1988 Mar 1;250(2):335-41. doi: 10.1042/bj2500335.
On the basis of the amino acid sequence of bovine rhodopsin, a series of peptides from the C-terminus (Rhod-4 and Rhod-1) and external loops (Rhod-10) were synthesized. Rabbit antisera to these peptides recognize the rhodopsin molecule in whole retina from 8-week-old normal and affected rcdl (rod/cone-dysplasic) Irish setters (8- and 4-weeks-old). When the rhodopsin content was equalized by using a solid-phase radioimmunoassay, the reaction with anti-peptide antisera to the C-terminal octapeptide (residues 341-348) is severely decreased in the rcdl-dog retinas. The results of mixing experiments suggest that this is not due to proteolytic clipping of the rhodopsin C-terminus from the affected dogs. Treatment of retinas with 1.0 mM-NaF, a phosphatase inhibitor, or pretreatment with alkaline and acid phosphatases does alter the reaction of the rhodopsin with anti-rhodopsin antisera. This suggests that the decreased reaction of the affected rhodopsin with the anti-peptide antisera may partially result from differences in intrinsic rhodopsin phosphorylation. However, since the reaction of rcdl retinas cannot be restored to that of the normals, these results suggest that the rhodopsin molecule from the rcdl dogs may be structurally altered in other ways.
基于牛视紫红质的氨基酸序列,合成了一系列来自C端(Rhod - 4和Rhod - 1)和外环(Rhod - 10)的肽段。针对这些肽段的兔抗血清可识别8周龄正常和患rcdl(视杆/视锥发育不良)的爱尔兰赛特犬(8周龄和4周龄)整个视网膜中的视紫红质分子。当使用固相放射免疫测定法使视紫红质含量相等时,在rcdl犬视网膜中,与针对C端八肽(残基341 - 348)的抗肽抗血清的反应严重降低。混合实验结果表明,这并非由于患犬视紫红质C端的蛋白水解剪切所致。用1.0 mM - NaF(一种磷酸酶抑制剂)处理视网膜,或先用碱性和酸性磷酸酶预处理,确实会改变视紫红质与抗视紫红质抗血清的反应。这表明,患犬视紫红质与抗肽抗血清反应降低可能部分是由于视紫红质内在磷酸化的差异。然而,由于rcdl视网膜的反应无法恢复到正常水平,这些结果表明,rcdl犬的视紫红质分子可能在其他方面发生了结构改变。